论文部分内容阅读
克隆测定了传染性喉气管炎病毒(ILTV)北京E3和Zhonghai毒株的糖蛋白G基因(gG)序列,并与多种动物的Ⅰ型α-疱疹病毒gG基因序列作了比对.通过克隆gG基因至原核表达载体pET30a(+)构建pET30a-gG质粒,IPTG诱导表达含pET30a-gG重组质粒的阳性宿主菌后,分离纯化得到分子量约35kD的His-GG融合蛋白.利用该融合蛋白制备的纯化的抗His-GG大鼠多抗血清,观测ILTV糖蛋白G的在细胞上的分布,发现糖蛋白G主要存在于宿主细胞的细胞核外周及细胞膜上,在散在细胞内的分布较为分散,而在融合细胞的结合部位分布较为集中.利用抗体中和病毒糖蛋白G的实验结果表明,糖蛋白G可能参与病毒从细胞到细胞的直接感染(CTCS),从而影响病毒复制和噬斑的形成.
The glycoprotein G gene (gG) sequence of infectious laryngotracheitis virus (ILTV) from Beijing E3 and Zhonghai strains was cloned and compared with the gG gene sequence of a type of α-herpesvirus of various animals. gG gene to prokaryotic expression vector pET30a-gG plasmid, IPTG induced expression of recombinant plasmid containing pET30a-gG positive host bacteria, isolated and purified to give a molecular weight of about 35kD His-GG fusion protein prepared using the fusion protein The purified antiserum against His-GG rats was used to observe the distribution of ILTV glycoprotein G on the cells. It was found that glycoprotein G mainly exists on the periphery of the nucleus and on the cell membrane of the host cells, and is more dispersed in scattered cells. The results showed that glycoprotein G might be involved in the direct cell-to-cell infection (CTCS) and thus affect viral replication and plaque formation.